Antibiotics are important adjuncts to oral rehydration therapy in cholera disease management. However, due to the rapid emergence of resistance to the antibiotics used to treat cholera, therapeutic options are becoming limited. This problem is more pronounced when treating children, where the use of many of the preferred antibiotics is limited by age-related contraindications. There is, therefore, a critical need to develop additiona therapeutics to aid in the treatment of cholera. The goal of this application is to identify lead compounds that will be developed further to complement existing cholera treatments, particularly for children who are at greatest risk from severe dehydration. The objective of this proposal is to identify and characterize small molecule inhibitors of the extracytoplasmic stress response (sE) pathway of V. cholerae. These compounds will have the potential to be developed into therapeutic agents and will also be valuable probes for uncovering basic molecular mechanisms of an important cause of diarrheal disease. Guided by strong preliminary data, the objective of this proposal will be accomplished by pursuing three specific aims: 1) Identify and validate small molecule inhibitors of sE activity using high-throughput screening of a chemical library; 2) Determine the mechanism of action of validated inhibitors; and 3) Determine compound effectiveness in an animal model of cholera infection. At the successful completion of this project several lead compounds that inhibit this stress response pathway in V. cholerae will have been identified and characterized. Also, compounds that are effective in reducing V. cholerae colonization in an animal model of disease will have been identified. This contribution will be significant because it is the first step toward the development of new and fundamentally different therapeutics for cholera.